GSM6932009: C6,H3K27ac,Input; Homo sapiens; ChIP-Seq
CRC tissue was fixed in 1% formaldehyde for 10min at room temperature, after which 0.125M glycine was added and the mixture was sat for 5min to terminate the crosslinking reaction. The tissue was then treated with cell lysis buffer and nucleus was collected by centrifuging at 800g for 5min at 4 °C. Next, nucleus was treated with nucleus lysis buffer and sonicated to fragment chromatin DNA. Antibodies against H3K27ac (Abcam, USA) and a nonspecific rabbit IgG (Santa Cruz, USA) were incubated overnight with the crosslinked protein and DNA for immunoprecipitation reactions with protein A/G magnetic beads. DNA fragments were purified and collected by a Dr.GenTLE Precipitation Carrier kit (Takara, Japan). Libraries were prepared for sequencing using stranded Illumina protocols